In today's lab we evaluated the results that we obtained through sequencing. We had sent in two samples to be sequenced and were able to obtain some results from only one of the samples. Sample number 2 did have a small insert into the T-Vector, but most of the insert was labeled "N" which means uncertain nucleotide position. Since the insert contained only one "certain" nucleotide, no BLAST could be performed on this insert. Sample number 4 did also have a short insert sequence of 27bp that we entered into BLAST. No results were found using different parameters (non-human, non-mouse, nucleotide collection, and, NCBI genomes). We did have success in our BLAST when we used the protein data bank parameter, but the top results were from: Chain A, Crystal Structure Of The Hybrid State Of Ribosome In Complex With The Guanosine Triphosphatase Release Factor 3, which is not close to our gene of interest: OOMT2 gene in roses. Some error could have occurred because there were also a great number of "N" bases included in the 27bp fragment that was used in the BLAST.
This concludes our project. We were successful in obtaining DNA from our organism, as well as removing the intron from the gene, and rejoining our fragments, but were unsuccessful in ligating our gene into a T-Vector to proceed with the cloning.
Written by: Kim Lovik